HBV)微染色体重塑与病毒复制。方法:HepG2细胞单独转染、共转染线性HBV单体与PPARα表达或PPARα沉默载体;Southern blot检测转染细胞质内HBV核心颗粒DNA;实时定量PCR定量检测细胞质HBV核心颗粒DNA及细胞核内HBV共价闭合环状DNA(covalently closed circular DNA
Objective:To determine whether peroxisome proliferator-activated receptor alpha(PPARα)is involved in the regulation of hepatitis B virus(HBV)minichromosome remodeling and viral replication. Methods:Linear HBV DNA monomers or co-transfected PPARα over-expression or silencing vectors were transfected into HepG2 cells. HBV DNA from core particles was detected in Southern blot and nuclear HBV covalently closed circular DNA(cccDNA)was isolated from HBV-transfected cells
quantified by real-time PCR using the specific primers and probe. Heptitis B surface antigen(HBsAg)and Heptitis B e antigen(HBeAg)in HepG2 culture supernatants were measured by enzyme-linked immunosorbent assay(ELISA). And chromatin immunoprecipitation(Ch IP) was utilized to detect the recruitment of PPARα
histones and chromatinmodifying enzymes onto cccDNA in transfected HepG2 cells. Results:The PPARα over-expression enhanced the recruitment of PPARα onto cccDNA
increased the levels of HBV open circular DNA(OC DNA)and the secreted viral productions in the culture supernatant
e.g. HBsAg and HBeAg
and also contributed to the recruitment of acetylated H3 and H4 and acetyltransferase p300 onto HBV cccDNA compared with those levels in cells transfected with linear HBV DNA alone. Meanwhile
those levels in cells with co-transfection of PPARα sh RNA were opposite. Conclusion:PPARα directly bound to cccDNA and contributed to the recruitement of acetyltransferase p300 onto HBV cccDNA. The modulation of PPARα on HBV cccDNA epigenetics and the regulation on HBV replication. This findings provide new insights into the molecular mechanisms associated with HBV-host transcription factors interactions.